F. oxysporum f.sp. melonis race 1,2-resistant melons

Abstract

Methods for conveying Fusarium oxysporum f.sp. melonis (FOM) race 1,2 resistance into non-resistant melon germplasm are provided, in some embodiments, the methods include introgressing FOM race 1,2 resistance into a non-resistant melon using one or more nucleic acid markers for marker-assisted selection among melon lines to be used in a melon breeding program, wherein the markers are linked to FOM race 1,2 resistance. Also provided are quantitative trait loci (QTLs) associated with resistance to FOM race 1,2; isolated and purified genetic markers associated with FOM race 1,2 resistance; melon plants, seeds, and tissue cultures produced by any of the disclosed methods; fruit and seed produced by the disclosed melon plants; and compositions including amplification primer pairs capable of initiating DNA polymerization by a DNA polymerase on melon nucleic acid templates to generate melon marker amplicons.

What is claimed is: 
     
       1. A method for the production of an inbred melon plant adapted for conferring, in hybrid combination with a suitable second inbred, resistance to  F. oxysporum  f.sp.  melonis  (FOM) race 1,2, the method comprising:
 (a) selecting a first donor parental line possessing a desired FOM race 1,2 resistance and having a resistant locus mapping to linkage group 9 and mapped by one or more of the markers 9.1, 9.2, and 9.3; 
 (b) crossing the first donor parent line with a second parental line in hybrid combination to produce a segregating plant population; 
 (c) screening the segregating plant population for identified chromosomal loci of one or more genes associated with the resistance to FOM race 1,2; and 
 (d) selecting plants from the population having the identified chromosomal loci for further screening until a line is obtained which is homozygous for resistance to FOM race 1,2 at sufficient loci to give resistance to FOM race 1,2 in hybrid combination. 
 
     
     
       2. A method for producing melon plants which are resistant to  F. oxysporum  f.sp.  melonis  (FOM) race 1,2 occurring in melon, the method comprising:
 (e) providing a  Cucumis melo  plant which contains one or more alleles that confer resistance to FOM race 1,2, which alleles are present within one or more of five Quantitative Trait Loci QTL1-QTL5 on different chromosomes, wherein:
 (i) QTL1 is defined by the following markers:
 (1) a marker of about 322 basepairs (bp), wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 1 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 2; and 
 (2) a marker of about 303 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 3 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 4; 
 or any part of a DNA sequence as in 03MFR001795 linked within 1, 2, 5, or 10 cM to at least one of the markers of (1) and (2) conferring resistance to FOM race 1,2; 
 
 (ii) QTL2 is defined by the following markers:
 (1) a marker of about 237 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 5 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 6; and 
 (2) a marker of about 189 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 7 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 8, 
 or any part of a DNA sequence as in 03MFR001795 linked within 1, 2, 5, or 10 cM to at least one of the markers of (1) and (2) conferring resistance to FOM race 1,2; 
 
 (iii) QTL3 is defined by the following markers:
 (1) a marker of about 221 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 9 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 10; and 
 (2) a marker of about 229 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 11 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 12, 
 or any part of a DNA sequence as in 03MFR001795 linked within 1, 2, 5, or 10 cM to at least one of the markers of (1), and (2) conferring resistance to FOM race 1,2; 
 
 (iv) QTL4 is defined by the following markers:
 (1) a marker of about 329 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 13 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 14; 
 (2) a marker of about 279 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 15 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 16; and 
 (3) a marker of about 147 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 17 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 18, 
 or any part of a DNA sequence as in 03MFR001795 linked within 1, 2, 5, or 10 cM to at least one of the markers of (1), (2), and (3) conferring resistance to FOM race 1,2; and 
 
 (v) QTL5 is defined by the following markers:
 (1) a marker of about 246 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 19 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 20; and 
 (2) a marker of about 251 bp, wherein the marker corresponds to an amplification product generated by amplifying a  Cucumis melo  nucleic acid with a forward primer comprising a nucleotide sequence as set forth in SEQ ID NO: 21 and a reverse primer comprising a nucleotide sequence as set forth in SEQ ID NO: 22, 
 or any part of a DNA sequence as in 03MFR001795 linked within 1, 2, 5, or 10 cM to at least one of the markers of (1) and (2) conferring resistance to FOM race 1,2; and 
 
 (f) crossing the  Cucumis melo  plant provided in step (a) with  Cucumis melo  culture breeding material to produce one or more progeny individuals, 
 
 whereby one or more melon plants which are resistant to  F. oxysporum  f.sp.  melonis  (FOM) race 1,2 occurring in melon are produced. 
 
     
     
       3. The method of  claim 2 , further comprising:
 (a) collecting the seeds resulting from the cross in step (b); 
 (b) regenerating the seeds into plants; 
 (c) evaluating the plants of step (d) for resistance to FOM race 1,2; and 
 (d) identifying and selecting plants which are resistant to the FOM race 1,2. 
 
     
     
       4. The method of  claim 2 , wherein the  Cucumis melo  plant provided in step (a) is 03MFR001795 or an ancestor or descendent thereof. 
     
     
       5. A method for identifying a first  Cucumis melo  plant or germplasm that displays resistance, improved resistance, or reduced susceptibility to FOM race 1,2, the method comprising detecting in the first  Cucumis melo  plant or germplasm at least one allele of one or more marker locus that is associated with the tolerance, improved tolerance, reduced susceptibility, or susceptibility, wherein the one or more marker locus is selected from the group consisting of: (a) 9.1, 9.2, 9.3; (b) a marker locus linked to a marker locus of (a); and (c) a marker locus localizing within a chromosome interval including a marker pair 9.1 and 9.3. 
     
     
       6. The method  claim 5 , wherein the detecting comprises detecting at least one allelic form of a polymorphic simple sequence repeat (SSR) or a single nucleotide polymorphism (SNP). 
     
     
       7. The method  claim 5 , wherein the detecting comprises amplifying the marker locus or a portion of the marker locus and detecting the resulting amplified marker amplicon. 
     
     
       8. The method of  claim 7 , wherein the amplifying comprises employing a polymerase chain reaction (PCR) or ligase chain reaction (LCR) using a nucleic acid isolated from the first melon plant or germplasm as a template in the PCR or LCR. 
     
     
       9. The method  claim 5 , wherein the at least one allele is a favorable allele that positively correlates with FOM race 1,2 resistance or improved FOM race 1,2 resistance.